INFLUÊNCIA DE POLISSACARÍDEOS SULFATADOS SOBRE A MOTILIDADE E MORFOLOGIA ESPERMÁTICA PÓS DESCONGELAÇÃO DE PROCHILODUS BREVIS

This research aims to verify the influence of sulfated polysaccharides, extracted from the skin of tilapia, on the after thawing motility and morphology of P. brevis sperm. For this, 17 males were hormonally induced to reproduce, through the application of two doses of pituitary carp extract, 0.4 and 4.0mg kg-1. After the seminal collection, objective analyzes were performed and samples with motility greater than 80% were selected to form the pools. Then, the pools were frozen in solution supplemented, or not, with different concentrations of glycosaminoglycans (GAGs): 0 (control); 0.5; 1.0; 1.5; 2.0; 2.5; 3.0; 3.5; 4.0; 4.5 or 5.0mg mL-1 (total of 10 treatments). The samples were placed in 0.25 mL French straws and submitted to an equilibrium time of 10 minutes at 4 oC. Then, they were kept in the dry shipper for 15 minutes and finally stored in liquid nitrogen. After 15 days, samples were thawed in a water bath at 30 oC for 16 seconds and evaluated for sperm motility and morphology. Thus, it was observed that the increase in GAGs caused a decrease in sperm motility, however the control and the concentration of 0.5mg mL-1 presented very similar data. On the other hand, no decrease in normal sperm was observed with an increase in the concentration of GAGs. Therefore, it is concluded that the lowest concentration of GAGs is the most adequate to supplement the sperm freezing medium.Esta pesquisa tem como objetivo verificar a influência de polissacarídeos sulfatados, extraídos da pele da tilápia, na motilidade e morfologia pós-descongelamento de espermatozoides de P. brevis. Para isso, 17 machos foram induzidos hormonalmente à reprodução, através da aplicação de duas doses de extrato de pituitária de carpa, 0,4 e 4,0mg kg-1. Após a coleta seminal, análises objetivas foram realizadas e amostras com motilidade superior a 80% foram selecionadas para formar os pools. Em seguida, os pools foram congelados em solução suplementada, ou não, com diferentes concentrações de glicosaminoglicanos (GAGs): 0 (controle); 0,5; 1,0; 1,5; 2,0; 2,5; 3,0; 3,5; 4,0; 4,5 ou 5,0mg mL-1 (total de 10 tratamentos). As amostras foram colocadas em palhetas francesas de 0,25 mL e submetidas a um tempo de equilíbrio de 10 minutos a 4 oC. Em seguida, foram mantidos no dry shipper por 15 minutos e, por fim, armazenados em nitrogênio líquido. Após 15 dias, as amostras foram descongeladas em banho-maria a 30 oC por 16 segundos e avaliadas quanto à motilidade e morfologia espermática. Assim, observou-se que o aumento dos GAGs causou diminuição da motilidade espermática, porém o controle e a concentração de 0,5mg mL-1 apresentaram dados muito semelhantes. Por outro lado, não foi observada diminuição de espermatozóides normais com o aumento da concentração de GAGs. Portanto, conclui-se que a menor concentração de GAGs é a mais adequada para complementar o meio de congelamento de espermatozoides

Effects of a sulfated polysaccharide isolated from the red seaweed Solieria filiformis on models of nociception and inflammation

AbstractThis work reports the effects of a sulfated polysaccharide (SP-Sf), isolated from the seaweed Solieria filiformis and characterized by Fourier transformed infrared (FT-IR), on nociception and inflammation. Male Swiss mice were pretreated with SP-Sf 30min before receiving an injection of 0.8% acetic acid, 1% formalin or 30min prior to a thermal stimulus. We observed that SP-Sf (1, 3 or 9mg/kg) significantly reduced the number of writhes. SP-Sf also reduced the second phase of the formalin test and did not cause a significant antinociceptive effect in the hot plate test, suggesting that its antinociceptive action occurs through a peripheral mechanism. SP-Sf (1, 3 or 9mg/kg) did not show a significant anti-inflammatory effect in Wistar rats when administrated by the systemic route 1h before testing using carrageenan or dextran. Finally, SP-Sf (9mg/kg) did not show significant signs of toxicity when administrated in mice

Carragenana da epífita Hypnea musciformis obtida do cultivo experimental de Solieria filiformis em Flecheiras, Estado do Ceará, Brasil = Carragennan from Hypnea musciformis epiphyte obtained from Solieria filiformis experimental culture at Flecheiras, Ceará State, Brazil

O cultivo da rodoficea Solieria filiformis vem sendo desenvolvido na praia de Flecheiras, Estado do Ceara, como perspectiva de utilizacao sustentavel do recurso para biotecnologia, mas o crescimento de epifitas sobre a especie compromete seu bom crescimento. Desta forma, avaliou-se a eficiencia de duas metodologias (M I e M II) noisolamento da Kappa-carragenana (ƒÈ-CAR) da alga marinha vermelha Hypnea musciformis. A ƒÈ-CAR foi obtida mediante digestao proteolitica (M I) ou agua quente (80‹C) (M II). Os extratos foram analisados quanto a sua composicao quimica, seguido por cromatografia de troca ionica em coluna de DEAE-celulose usando um gradiente de NaCl. A atividade anticoagulante foi avaliada pelo teste do tempo de tromboplastina parcial ativada (TTPA), usando plasma humano normal e heparina (HEP) padrao (193 UI mg-1). A diferenca nos rendimentos foi 4,28 } 0,26%% (p Solieria filiformis (Rhodophyceae) culture is being developed at Flecheiras Beach, state of Ceara, Brazil, for its future sustainable use in biotechnology. However, epiphyte growth on the species jeopardizes suitable development. The efficiency of two methods (M I and M II) in the isolation of kappa-carrageenan (ƒÈ-CAR) from red marine alga Hypnea musciformis has been evaluated. ƒÈ-CAR was obtained by proteolytic digestion (M I) or by hot water (80‹C)(M II) and extracts were analyzed with regard to their chemical composition and then by ion-exchange chromatography on DEAE-cellulose column with NaCl gradient. Anticoagulant activity was evaluated by the activated partial thromboplastine time (APTT) test with common human plasma and standard heparin (HEP) (193 IU mg-1). Yield difference reached 4.28 } 0.26%% (p < 0.05), and the lowest sulfate content was obtained with M I (17.00%) when compared to that with M II (26.30%). Further, different chromatographic profiles were reported between the two methods. Whereas M I was efficacious in obtaining ƒÈ-CAR anticoagulants (2.67 IU mg-1), alternative M II was efficient for the food industry. Results suggest that ƒÈ-CAR extraction method from H. musciformis depends on the industryfs usage criterion

Carragenana da epífita Hypnea musciformis obtida do cultivo experimental de Solieria filiformis em Flecheiras, Estado do Ceará, Brasil - doi: 10.4025/actascitechnol.v33i2.9096

Solieria filiformis (Rhodophyceae) culture is being developed at Flecheiras Beach, state of Ceará, Brazil, for its future sustainable use in biotechnology. However, epiphyte growth on the species jeopardizes suitable development. The efficiency of two methods (M I and M II) in the isolation of kappa-carrageenan (κ-CAR) from red marine alga Hypnea musciformis has been evaluated. κ-CAR was obtained by proteolytic digestion (M I) or by hot water (80°C) (M II) and extracts were analyzed with regard to their chemical composition and then by ion-exchange chromatography on DEAE-cellulose column with NaCl gradient. Anticoagulant activity was evaluated by the activated partial thromboplastine time (APTT) test with common human plasma and standard heparin (HEP) (193 IU mg-1). Yield difference reached 4.28 ± 0.26%% (p < 0.05), and the lowest sulfate content was obtained with M I (17.00%) when compared to that with M II (26.30%). Further, different chromatographic profiles were reported between the two methods. Whereas M I was efficacious in obtaining κ-CAR anticoagulants (2.67 IU mg-1), alternative M II was efficient for the food industry. Results suggest that κ-CAR extraction method from H. musciformis depends on the industry’s usage criterion.O cultivo da rodofícea Solieria filiformis vem sendo desenvolvido na praia de Flecheiras, Estado do Ceará, como perspectiva de utilização sustentável do recurso para biotecnologia, mas o crescimento de epífitas sobre a espécie compromete seu bom crescimento. Desta forma, avaliou-se a eficiência de duas metodologias (M I e M II) no isolamento da Kappa-carragenana (κ-CAR) da alga marinha vermelha Hypnea musciformis. A κ-CAR foi obtida mediante digestão proteolítica (M I) ou água quente (80°C) (M II). Os extratos foram analisados quanto à sua composição química, seguido por cromatografia de troca iônica em coluna de DEAE-celulose usando um gradiente de NaCl. A atividade anticoagulante foi avaliada pelo teste do tempo de tromboplastina parcial ativada (TTPA), usando plasma humano normal e heparina (HEP) padrão (193 UI mg-1). A diferença nos rendimentos foi 4,28 ± 0,26%% (p < 0,05). O menor teor de sulfato foi obtido no M I (17,00%) quando comparado ao M II (26,30%) e diferentes perfis cromatográficos foram observados entre os métodos. O M I demonstrou ser eficiente na obtenção de κ-CAR anticoagulantes (2,67 UI mg-1), enquanto o M II é alternativo para indústria de alimentos. Os resultados sugerem que o método de obtenção da κ-CAR da rodofícea H. musciformis a ser empregado depende do critério de utilização na indústria

Iota-carrageenans from Solieria filiformis (Rhodophyta) and their effects in the inflammation and coagulation=Iota-carragenanas da rodofícea Solieria filiformis e seus efeitos na inflamação e coagulação

Biochemical analyses are important tools for discovering new bioactive compounds for medical clinic. This study aimed at isolating iota-carrageenans (ι-CARs) from Solieria filiformis (Rhodophyta) by enzymatic extraction (EE), refined hot-water extraction (RHWE) and hot-water extraction (HWE), and test (EE, s.c.) their anti-inflammatory effects in the peritonitis model using the Lambda-carrageenan (700 µg cavity-1, i.p.) as an inflammatory stimuli in rats. The activated partial thromboplastin time (APTT) was also evaluated in ι-CARs fractions, obtained by ion-exchange chromatography (DEAE-cellulose), using rabbit plasma and compared to heparin (193 IU mg-1). The results showed that the ι-CARs (EE) (3 or 9 mg kg-1) containing 89.92% total sugars, 29.02% sulfate and absence of contaminant proteins inhibited (p 0.05). Similar chromatography profiles were obtained among the methods; however, with fractions revealing different pattern on charge density by electrophoresis. Fractions had no virtually effects on APTT (1.16, 1.73 and 1.59 IU mg-1 for EE, RHWE and HWE, respectively). Further investigations to better understanding the actions of S. filiformis ι-CARs (EE) in the inflammatory response are suggested.Análises bioquímicas são ferramentas importantes para a descoberta de novos compostos bioativos para clínica médica. Neste estudo, iota-carragenanas (ι-CARs) da rodofícea Solieria filiformis foram isoladas (extração enzimática (EE), extração aquosa a quente refinada (EAQR) ou extração aquosa a quente (EAQ)) e testadas (EE; s.c.) no modelo de peritonite, usando-se a Lambda-carragenana (700 µg cavidade-1; i.p.) como um estímulo flogístico, para averiguar seus efeitos anti-inflamatórios em ratos. O tempo de tromboplastina parcial ativada (TTPA) também foi avaliado em frações de ι-CARs, obtidas por cromatografia de troca iônica (DEAE-celulose), usando plasma de coelho e comparadas à heparina (193 UI mg-1). Verificaram-se que ι-CARs (EE) (3 ou 9 mg kg-1) contendo 89,92% de açúcares totais, 29,02% de sulfato e destituídas de contaminação proteica, inibiram (p 0,05). Os perfis cromatográficos mostraram-se semelhantes entre os métodos de extração, porém revelando, por eletroforese, frações com diferenças em termos de densidade de carga. As frações praticamente não alteraram o TTPA (1,16; 1,73 e 1,59 UI mg-1 para EE, EAQR e EAQ, respectivamente). Investigações são sugeridas para melhor compreender as ações das ι-CARs (EE) da S. filiformis na resposta inflamatória

In vivo toxicological evaluation of crude sulfated polysaccharide from the green seaweed Caulerpa cupressoides var. lycopodium in Swiss mice - doi: 10.4025/actascitechnol.v35i4.15699

Seaweeds are widely consumed as vegetables and medicinal products. The green seaweed Caulerpa cupressoides var. lycopodium sulfated polysaccharides (SPs) demonstrated anticoagulant (in vitro) and anti- and prothrombotic (in vivo) effects. However, their toxicity in vivo has not been fully determined. This study evaluated their toxicity in vivo in male Swiss mice. Animals (20-26 g, six group-1) received crude SP (9 mg kg-1, i.p.) or 0.9% saline (0.1 mL 10 g-1, i.p.) for 14 consecutive days, and then analyzed the wet weight of animal’s body organs-1 and biochemical/hematological parameters. Histopathological evaluation was also performed related to crude SP treatment. The results showed that crude SP did not cause toxicity and mortality. Regarding the biochemical analyses, crude SP did not lead to hepatic or renal dysfunctions, but affected (p < 0.05) the platelet count (1530.75 ± 1.05 × 103 µL-1) compared with the control (969.75 ± 0.51 × 103 µL-1) according to the hematological evaluation. Although histological changes in the liver and kidney have occurred, results suggested reversibility. The increased spleen size (p < 0.05) also had no toxicological significance based on histopathological analysis. Therefore, crude SP from C. cupressoides could represent safe pharmacological tool in future studies on immunomodulation and thrombosis in vivo.